From Membrane to Nucleus: T eaching Cell S ignaling via the R as Pathway

1. From Membrane to Nucleus: Teaching Cell Signaling via the Ras Pathway

2. Cell Biology Team Kathleen Marrs, IUPUI James Marrs, IUPUI Mariah Judd, IUPUI Todd Murphy, Ivy Tech Community College (IUPUI) Robert Steven, University of Toledo TomerAvidor-Reiss, University of Toledo Henry Chang, Purdue University Facilitators: Brian Kinkle, University of Cincinnati Sadie Hebert, University of Minnesota

3. Context • This unit was prepared for a sophomore level cell biology course, taught mid way through the semester • This unit was designed with the flexibility to be scaled down to a freshman level biology course. • Unit designed to take 1-2 (one hour) class sessions

4. Expected Background • Basics of cell structure and organelles • Protein control by phosphorylation • Understand that cells have the ability to respond to extracellular stimuli • Basics of cell signaling cascades (including various types of G-proteins)

5. Teachable Unit Goals and Outcomes • Know specific components and relationship between components of the Ras pathway affecting cell proliferation. • Order the sequence of proteins involved in the Ras pathway and describe their cellular location (BT 1-2) • Describe the role of each component of the Ras pathway (BT 2-3) • Evaluate the consequences of perturbations to the signaling pathway using real data. • Analyze the effect of mutations on the components of the Ras pathway (BT 5-6) • Practice the skill of analyzing data, controls and outcomes of experiments. (BT 4-5)

6. Teachable Unit Goals and Outcomes • Know specific components and relationship between components of the Ras pathway affecting cell proliferation. • Create a schematic depicting the RTK/RAS/MAP kinase signaling pathway in both the active and inactive states. (BT 1-2) • Discuss the role and significance of each component of the Ras pathway. (BT 2-3) • Evaluate the consequences of perturbations to the signaling pathway using real data. • Analyze the effect of perturbation(s) on the components of the Ras pathway. (BT 4-5) • Develop the skill of analyzing data, controls and outcomes of experiments. (BT 4-5) (BT=Bloom’s Taxonomy)

7. Pre-Assignments • Read the introduction from “The History of Ras: the Saga Continues” by Cox and Der, 2010 • Review chapter section in text book that covers Ras pathway and review section questions • MAPK pathway video by Genentech (Youtube video) with video reflections

8. Pre-Assignments • Read the introduction from “The History of Ras: the Saga Continues” by Cox and Der, 2010 • Review chapter section in text book that covers Ras pathway and review section questions • MAPK pathway video by Genentech (Youtube video) with video reflections

9. Pre-Assignments Summary Modulation of signalling by Sprouty: a developing story Hong Joo Kim & Dafna Bar-Sagi Nature Reviews Molecular Cell Biology 5, 441-450 (June 2004) doi:10.1038/nrm1400

10. From Membrane to Nucleus: Our Tidbit Teaching Cell Signaling via the Ras Pathway

11. Activation of a G-protein involves ___________. • Cleavage of the G-protein • Methylation of the G-protein • Binding of GTP • Binding of GDP • Techno music Clicker Question

12. Ras is a __________. • Signal transduction molecule • Transcription factor • Calcium channel • Membrane receptor • Flavor of the Month at Starbucks Clicker Question

13. Why Do We Care About Ras? • Ras is the most common oncogene in human cancer. • 90% of all pancreatic cancers • 25% of all human tumors • The Ras pathway provides a focus on the origins of cancer at the molecular level. • Ras is a key cellular protein for cell growth, differentiation and survival. http://www.sciencephoto.com/

14. Ras-zle Dazzle Puzzle Ras pathway modeling activity Materials needed: • One 8.5x11 puzzle board • 2 Envelopes labeled #1 “OFF” and #2 “ON” filled with puzzle pieces Working with your pod… • First: read and open envelope #1 “OFF” and assemble the pieces in the appropriate place on the puzzle board. (5 minutes) • Second: share your puzzle with the other pods at your table to compare and discuss. (5 minutes)

15. Ras-zle Dazzle Puzzle “OFF” State

16. Ras-zle Dazzle Puzzle Ras pathway modeling activity Working with your pod… • Third: read and open envelope #2 “ON” and add the new pieces to the puzzle board. (5 minutes) • Fourth: share your completed puzzle with the other pods at your table to compare and discuss. (5 minutes)

17. Ras-zle Dazzle Puzzle “ON” State

18. What initially triggers a change in gene expression (in a WT cell)? • Ras Activation • GTP Hydrolysis • MAP Kinase Phosphorylation • Growth Factor – Receptor binding Clicker Question

19. Ras-zle Dazzle Puzzle:What is the Significance? • What is the significance and outcome of… • the receptor – ligand interaction • the Ras activation • the MAP kinase cascade • What is the significance of the ERK translocation and what is one possible outcome?

20. When Good Cells Go Bad:Disruptions in the pathway • What happens when we perturb the pathway? Summative Assessment

21. Summative homework • Practice with data analysis, controls and outcomes. • Uses real data from primary literature. Summative Assessment

22. Rita Levi-Montalcini “Above all, don’t fear difficult moments. The best comes from them.” In Praise of Imperfection, Levi-Montalcini’s biography • Nobel laureate • Discovered growth factor signaling • Italian Jewish scientist • Lived to age 103 Image

23. TidbitGoals and Outcomes Goal I Know specific components and relationship between components of the Ras pathway affecting cell Create a schematic depicting the RTK/RAS/MAP kinase signaling pathway in both the active and inactive states.(BT 1-2) Discuss the role and significance of each component of the Ras pathway(BT 2-3) Outcome a) Outcome b)

24. With thanks to HHMI, The University of Minnesotaand the National Academies NorthStar (Midwest) Summer Institute (NANSI): Thanks also to Brian and Sadie, our amazing facilitators